Antibodies to human interleukin-2 induced by synthetic polypeptides

Abstract

Chemically synthesized polypeptides containing about 6 to about 40 amino acid residues having amino acid residue sequences that substantially correspond to the amino acid residue sequences of antigenic determinants of interleukin-2, when administered alone or as polymers or as conjugates bound to carriers, induce the production of antibodies of predetermined specificities. The polypeptides and the antibodies produced thereto can be used in diagnostic systems to measure the presence and amount of interleukin-2 and interleukin-2 cell surface receptors or binding sites in an assayed sample.

What is claimed is: 
     
       1. A synthetic polypeptide containing about 6 to about 40 amino acid residues having an amino acid residue sequence that substantially corresponds to an amino acid residue sequence of interleukin-2 that is selected from the group consisting of: (a) the interleukin-2 protein from about position 18 to about position 32 from the amino-terminus thereof,   (b) the interleukin-2 protein from about position 79 to about position 92 from the amino-terminus thereof,   (c) the interleukin-2 protein from about position 111 to about position 125 from the amino-terminus thereof, and   (d) the interleukin-2 protein from about position 139 to about position 153 from the amino-terminus thereof;   said synthetic polypeptide having the capacity alone, as a polymer or as a conjugate of said polypeptide bound to a carrier, when injected into a host in an effective amount and in a physiologically tolerable diluent of inducing the production of antibodies to interleukin-2.   
     
     
       2. The synthetic polypeptide according to claim 1 wherein said synthetic polypeptide contains from about 10 to about 20 amino acid residues. 
     
     
       3. The synthetic polypeptide according to claim 1 including the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula and selected from the group consisting of: ThrAsnSerAlaProThrSerSerSerThrLysLysThrGlnLeu,   LeuGluGluGluLeuLysProLeuGluGluValLeuAsnLeu,   ValIleValLeuGluLeuLysGlySerGluThrThrPheMetCys;and   AsnArgTrpIleThrPheCysGlnSerIleIleSerThrLeuThr.   
     
     
       4. The synthetic polypeptide according to claim 1 including the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: ThrAsnSerAlaProThrSerSerSerThrLysLysThrGlnLeu.   
     
     
       5. The synthetic polypeptide according to claim 1 including the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: LeuGluGluGluLeuLysProLeuGluGluValLeuAsnLeu.   
     
     
       6. The synthetic polypeptide according to claim 1 including the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: ValIleValLeuGluLeuLysGlySerGluThrThrPheMetCys.   
     
     
       7. The synthetic polypeptide according to claim 1 including the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: AsnArgTrpIleThrPheCysGlnSerIleIleSerThrLeuThr.   
     
     
       8. The synthetic polypeptide according to claim 1 wherein said physiologically tolerable diluent is a member of the group consisting of water, saline and an adjuvant. 
     
     
       9. The synthetic polypeptide according to claim 1 wherein said carrier is selected from the group consisting of keyhole limpet hemocyanin, keyhole limpet hemocyanin in incomplete Freund's adjuvant, alum, keyhole limpet hemocyanin-alum absorbed, keyhole limpet hemocyanin-alum absorbed-pertussis, edestin, thyroglobulin, tetanus toxoid and tetanus toxoid in incomplete Freund's adjuvant. 
     
     
       10. An inoculum comprising an effective amount of synthetic polypeptide containing about 6 to about 40 amino acid residues having an amino acid residue sequence that immunologically corresponds substantially to an amino acid residue sequence of interleukin-2 that is selected from the group consisting of: (a) the interleukin-2 protein from about position 18 to about position 32 from the amino-terminus thereof,   (b) the interleukin-2 protein from about position 79 to about position 92 from the amino-terminus thereof,   (c) the interleukin-2 protein from about position 111 to about position 125 from the amino-terminus thereof, and   (d) the interleukin-2 protein from about position 139 to about position 153 from the amino-terminus thereof;   and a physiologically tolerable diluent, said inoculum, when introduced into a host, being capable of inducing the production of antibodies in the host that immunoreact with interleukin-2.   
     
     
       11. The inoculum according to claim 10 wherein said synthetic polypeptide contains from about 10 to about 20 amino acid residues. 
     
     
       12. The inoculum according to claim 10 wherein said synthetic polypeptide includes the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula and selected from the group consisting of: ThrAsnSerAlaProThrSerSerSerThrLysLysThrGlnLeu,   LeuGluGluGluLeuLysProLeuGluGluValLeuAsnLeu,   ValIleValLeuGluLeuLysGlySerGluThrThrPheMetCys;and   AsnArgTrpIleThrPheCysGlnSerIleIleSerThrLeuThr.   
     
     
       13. The inoculum according to claim 10 wherein said synthetic polypeptide includes the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: ThrAsnSerAlaProThrSerSerSerThrLysLysThrGlnLeu.   
     
     
       14. The inoculum according to claim 10 wherein said synthetic polypeptide includes the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: LeuGluGluGluLeuLysProLeuGluGluValLeuAsnLeu.   
     
     
       15. The inoculum according to claim 10 wherein said synthetic polypeptide includes the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: ValIleValLeuGluLeuLysGlySerGluThrThrPheMetCys.   
     
     
       16. The inoculum according to claim 10 wherein said synthetic polypeptide includes the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula: AsnArgTrpIleThrPheCysGlnSerIleIleSerThrLeuThr.   
     
     
       17. The inoculum according to claim 10 wherein said physiologically tolerable diluent is a member of the group consisting of water, saline and an adjuvant claim 10. 
     
     
       18. The inoculum according to wherein said synthetic polypeptide is bound to a carrier. 
     
     
       19. The inoculum according to claim 18 wherein said carrier is selected from the group consisting of keyhole limpet hemocyanin, keyhole limpet hemocyanin in incomplete Freund's adjuvant, alum, keyhole limpet hemocyanin-alum absorbed, keyhole limpet hemocyanin-alum absorbed-pertussis, edestin, thyroglobulin, tetanus toxoid and tetanus toxoid in incomplete Freund's adjuvant. 
     
     
       20. Antibodies raised in an animal host to a synthetic polypeptide containing about 6 to about 40 amino acid sequence that immunologically corresponds substantially to an amino acid residue sequence of interleukin-2 that is selected from the group selected from: (a) the interleukin-2 protein from about position 18 to about position 32 from the amino-terminus thereof,   (b) the interleukin-2 protein from about position 79 to about position 92 from the amino-terminus thereof,   (c) the interleukin-2 protein from about position 111 to about 125 from the amino-terminus thereof, and   (d) the interleukin-2 protein from about position 139 to about position 153 from the amino terminus thereof;   said antibodies having the capacity to immunoreact with interleukin-2.   
     
     
       21. Antibodies according to claim 20 wherein said synthetic polypeptide includes a sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula and selected from the group consisting of: ThrAsnSerAlaProThrSerSerSerThrLysLysThrGlnLeu,   LeuGluGluGluLeuLysProLeuGluGluValLeuAsnLeu,   ValIleValLeuGluLeuLysGlySerGluThrThrPheMetCys;and   AsnArgTrpIleThrPheCysGlnSerIleIleSerThrLeuThr.   
     
     
       22. A diagnostic system for assaying for the presence of interleukin-2 comprising in biochemically active form antibodies raised in an animal host to a synthetic polypeptide containing about 6 to about 40 amino acid residues having an amino acid residue sequence that immunologically coresponds to at least a portion of interleukin-2 that is selected from the group consisting of: (a) the interleukin-2 protein from about positions 18 to about 32 from the amino-terminus thereof,   (b) the interleukin-2 protein from about position 79 to about position 92 from the amino-terminus thereof,   (c) the interleukin-2 protein from about position 111 to about position 125 from the amino-terminus thereof, and   (d) the interleukin-2 protein from about position 139 to about position 153 from the amino terminus thereof;   said antibodies immunoreacting with an admixed sample to be assayed that contains interleukin-2 to form an immunoreactant with interleukin-2, the presence of said immunoreactant being signalled by an indicating means.   
     
     
       23. The diagnostic system according to claim 22 wherein said synthetic polypeptides includes the sequence of amino acid residues, taken from left to right and in the direction from amino-terminus to carboxy-terminus, represented by the formula and selected from the group consisting of: ThrAsnSerAlaProThrSerSerSerThrLysLysThrGlnLeu,   LeuGluGluGluLeuLysProLeuGluGluValLeuAsnLeu,   ValIleValLeuGluLeuLysGlySerGluThrThrPheMetCys;and   AsnArgTrpIleThrPheCysGlnSerIleIleSerThrLeuThr.   
     
     
       24. The diagnostic system according to claim 22 further including an indicating means comprising enzyme-linked second antibodies, said second antibodies being raised to antibodies of the same class and from the same species as the first named antibodies, and signalling said immunoreaction by binding to said first named antibodies present in said immunoreactant, said signal being indicated by the reaction of said linked enzyme with an added substrate. 
     
     
       25. The diagnostic system according to claim 22 wherein said indicating means comprises a radioactive element bonded to said antibodies and said immunoreaction causes precipitation of said immunoreactant containing said radioactive element. 
     
     
       26. A diagnostic system for assaying for the presence of interleukin-2 in a body component comprising in separate containers (i) a first reagent that contains in biologically active form a synthetic polypeptide containing a sequence of about six to about forty amino acid residues that immunologically corresponds substantially to an amino acid residue sequence of interleukin-2 that is selected from the group consisting of:   (a) the interleukin-2 protein from about position 18 to about position 32 from the amino terminus thereof,   (b) the interleukin-2 protein from about position 79 to about position 92 from the amino terminus thereof,   (c) the interleukin-2 protein form about position 111 to about position 125 from the amino terminus thereof, and   (d) the interleukin-2 protein from about position 139 to about position 153 from the amino terminus thereof;   said polypeptide, when linked to a carrier as a conjugate and introduces in an effective amount as an inoculum into a host animal, being capable of inducing production of antibodies in the host that immunoreact with interleukin-2; and   (ii) a second reagent that contains in biochemically active form an anti-polypeptide antibody that immunoreacts with said synthetic polypeptide; along with a means for indicating the presence of an immunoreaction between said first and second reagents; said first and second reagents, when admixed in predetermined amounts in the presence of a predetermined amount of body component to be assayed, providing an amount of immunoreaction signalled by said indicating means, the amount of said immunoreaction being different from a known immunoreaction amount when interleukin-2 is not present in said body component.   
     
     
       27. A method for assaying for the presence of an antigenic determinant of interleukin-2 in a sample comprising: (a) providing an anti-polypeptide antibody to a synthetic polypeptide of claim 1;   (b) admixing a predetermined amount of said anti-polypeptide antibody with a predetermined amount of sample to be assayed for the presence of the antigenic determinant to which the anti-polypeptide antibody binds;   (c) maintaining that admixture for a period of time sufficient for said anti-polypeptide antibody to bind to the antigenic determinant present in the admixed sample; and   (d) determining the amount of binding between said anti-polypeptide antibody and said antigenic determinant of interleukin-2.   
     
     
       28. An affinity sorbant comprising an inert, solid support having linked thereto in biochemically active form antibodies raised in an animal host to a synthetic polypeptide containing about 6 to about 40 amino acid residues having an amino acid sequence that imaunologically corresponds substantially to an amino acid residue sequence of interleukin-2 that is selected from the group consisting of: (a) the interleukin-2 protein from about position 18 to about position 32 from the amino-terminus thereof,   (b) the interleukin-2 protein from about position 79 to about position 92 from the amino-terminus thereof,   (c) the interleukin-2 protein from about position 111 to about position 125 from the amino-terminus thereof, and   (d) the interleukin-2 protein from about position 139 to about position 153 from the amino-terminus there;   said affinity sorbant forming a reversible complex when admixed with an aqueous composition containing interleukin-2.   
     
     
       29. A synthetic polypeptide having an amino acid residue sequence, taken from left to right and in the direction of amino-terminus to carboxy-terminus, represented by the formula: ThrAsnSerAlaIroThrSerSerSerThrLysLy and the pharmaceutically acceptable salts thereof.     
     
     
       30. A synthetic polypeptide having an amino acid residue sequence, taken from left to right and in the direction of amino-terminus to carboxy-terminus, represented by the formula: LeuGluGluGluLeuLysProLeuGluGluValLeuAsnLe and the pharmaceutically acceptable salts thereof.   
     
     
       31. A sythetic polypeptide having an amino acid residue sequence, taken from left to right and in the direction of amino-terminus to carboxy-terminus, represented by the formula: ValIleValLeuGluLeuLysGlySerGluThrThrPh and the pharmaceutically acceptable salts thereof.     
     
     
       32. A synthetic polypeptide having an amino acid residue sequence, taken from left to right and in the direction of amino-terminus to carboxy-terminus, represented by the formula: AsnArgTrpIleThrPheCysGluSerIleIleSerTh and the pharmaceutically acceptable salts thereof.